摘要
目的:建立巢式逆转录聚合酶链反应( reverse transcriptase polymer ase chain reaction,RT PCR)扩增CK20mRNA,检测大肠癌淋巴结微转移。方法:据CK20 基因序列分别设计2 对引物,建立巢式RT- PCR 扩增体系,检测CK20 mRNA在30 例行根治性手术大肠癌患者187个淋巴结及10 例良性肠道疾病患者25 个淋巴结中的表达情况,将检测结果与传统组织学检查相比较。结果:大肠癌患者30 例187 个淋巴结有25例97个检测到CK20 mRNA表达,阳性率51 .87%(97/187),病理学检查仅发现18 例57 个淋巴结存在肿瘤转移;10例良性肠道疾病25 个淋巴结扩增结果为阴性。结论:巢式RT- PCR检测淋巴结CK20 mRNA的表达是诊断大肠癌微细胞转移灵敏特异的指标,可对精确大肠癌临床分期、判断患者预后及制定治疗方案提供一定的理论依据。
OBJECTIVE:To develop a nested reverse transcriptase polymerase chain reaction method to amplify cytokeratin 20 mRNA (CK20 mRNA) for detecting colorectal cancer micrometastases in lymph nodes and to evaluate its specificity and sensitivity.METHODS:The RT-PCR system was set up by using nest primer. The presence of CK20 mRNA was investigated by nested RT-PCR in 187 lymph nodes from 30 colorectal cancer patients who underwent curative operation. Tweenty-five lymph nodes from 10 patients with benign intestinal disease were served as negative control. The results of nested RT-PCR were compared with ones of routine histology.RESULTS:The positive rate of CK20 mRNA in 187 lymph nodes from 30 colorectalcancer patients was 51.87%(97/187), while only 57 samples were histologically positive.The expression of CK20 mRNA in 25 lymph nodes of 10 cases of the control group were all negative. CONCLUSIONS:Nested reverse transcriptasepolymerase chain reaction was a sensitive and specific method to detect the expression of CK20 mRNA in lymph nodes as a helpful marker of micrometastases of colorectal carcinoma cells. It is superior to routine histopathological assay in examing lymph node metastases. The detection of CK20mRNA in lymph nodes may contribute to improve the accuracy of clinical staging, determine patients' prognosis and provide information for adjuvant therapy.
出处
《肿瘤防治杂志》
2005年第5期348-352,共5页
China Journal of Cancer Prevention and Treatment
关键词
结肠直肠肿瘤/病理学
逆转录聚合酶链反应
角蛋白生物合成
淋巴转移
colorectal neoplasms/pathology
reverse transcriptase polymerase chain reaction
keratin/biosynthesis
lymphatic metastasis
